The graphene prepared by chemical vapor deposition onto copper foils was transferred onto a poly(ethylene terephthalate) (PET) soft substrate by the aid of poly-methyl methacrylate (PMMA).The soft G/AuNPs/GOD composite electrode based on PET substrate was fabricated using a protocol in which the uniform distribution of Au nanoparticles (AuNPs) was firstly obtained by controlling the evaporation of gold sol on a graphene surface, then thioglycolic acid (TGA) was modified on the AuNPs through Au-S bond, and finally glucose oxidase (GOD) was immobilized on the surface of AuNPs through acylation reaction between TGA and the GOD.Glucose was detected in the linear range from 0.05 to 10.55 mmol/L with a linear correlation coefficient (r) of 0.9955.The detection was performed in phosphate buffer solution (pH 7) at 25℃ with a working potential of 0.6 V (vs.SCE electrode), and the detection limit was 1 μmol/L (3σ).The G/AuNPs/GOD flexible electrode based on PET substrate provided a new pathway to detect glucose in special environments using wearable equipment, which enlarged the applied field of glucose detection.

Objective To study the expression of WWOX, ErbB2 and Ki67 protein in non-small cell lung cancer(NSCLC) and the relationships with cell proliferation. Methods WWOX and ErbB2 protein expressions were evaluated by immunohistochemistry in 81 NSCLC patients (50 squamous cell carcinomas,31 adenocarcinomas and 20 adjacent normal lung tissues) and the correlations with histopathologic cell proliferation were analyzed. Results WWOX expression was absent/reduced in 72.8% of NSCLC, while it was normal in 80.0% of adjacent normal lung tissues. WWOX expression was strongly associated with tumor histology, histologic grade and lymph node metastasis ( X2 = 5.44, P = 0.019 ; X2 = 4.740, P = 0.029, X2 = 4.51, P = 0.034 ), reduced WWOX expression was higher in squamous cell carcinomas and in poorly differentiated tumors. The overexpression of ErbB2 was associated with TNM stage and lymph node metastasis ( X2 = 6.90, P = 0.009 ; X2= 5.68, P=0.017). WWOX expression was negatively related to the overexpression of ErbB2 (r=-0.239, P <0.05 ). WWOX expression intensity was nega-tively related to the high index of cell proliferation (r=-0.252, P<0.05 ). Conclusion The loss of WWOX ex-pression plays different roles in tumorigenesis of NSCLC and is related to high aggressiveness of tumors. The loss of WWOX expression and the overexpression of ErbB2 can estimate the prognosis of NSCLC.

Objective:To explore the curative effects of individualized surgical method for treating pyriform sinus fistula (PSF) in children.Methods:A total of 43 PSF children treated in the Department of Pediatric Surgery of Henan Provincial People′s Hospital from June 2015 to November 2019 were selected, with 41 cases on the left side and 2 cases on the right side.There were 27 cases in the quiescent inflammatory infection stage and 16 cases in the acute inflammatory infection stage.According to patients′ condition, low-temperature plasma radiofrequency ablation, fistulectomy, abscess incision and drainage or combined operation were conducted.The postoperative complications and recurrence were observed.Results:Among 43 cases, there were 19 cases treated with low-temperature plasma radiofrequency ablation, 11 cases treated with low-temperature plasma radiofrequency ablation combined with abscess incision and drainage, 5 cases treated with low-temperature plasma radiofrequency ablation combined with fistulectomy, and 8 cases treated with fistulectomy.Four cases had postoperative hoarseness and recovered within 1-5 months.One case occurred pseudo healing after abscess incision and drainage, and healed after incision and debridement.Totally, 4 cases recurred, and the follow-up time was from 3 months to 57 months.Conclusions:Low-temperature plasma radiofrequency ablation has the advantages of minimally invasive, safe and effective, which provides a new treatment for PSF in children.Based on the condition of children, the choice of individualized treatment can significantly improve the curative effects.

Objective:To explore the strategy of selecting the second stage operation after neonatal enterostomy.Methods:Clinical data of 35 cases admitted to the Department of Pediatric Surgery of Henan Provincial People′s Hospital for enterostomy in neonatal period and radical operation in the second stage from May 2014 to December 2019 were subject to retrospective analysis.They were inquired about the medical history in detail before the operation, and received barium enema and colonoscopy, 24-hour delayed radiography, anorectal manometry and other examinations.During the operation, a multi-point intestinal biopsy and rapid frozen pathological examination were performed accor-ding to the individual′s condition, and the specific surgical method was determined in combination with intraoperative exploration.Postoperative observations were conducted for such complications as anastomotic leakage, intestinal obstruction, and abdominal infection, as well as such medium and long-term outcomes as intestinal function recovery, defecation, and nutritional status.Results:Nine cases were diagnosed with neonatal necrotizing enterocolitis (NEC) and received ostomy closure.Sixteen cases were diagnosed with NEC concomitant with intestinal stenosis and received the narrow bowel enterectomy and anastomosis combined with ostomy closure; 10 cases were diagnosed with Hirschsprung′s disease, including 5 cases of total colonic aganglionosis treated with Soave radical operation, 3 cases of long-segment Hirschsprung′s disease treated with subtotal colectomy, inverted ascending coloanal anastomosis with Deloyers method and second enterostomy, and 2 cases of normal-segment Hirschsprung′s disease treated with pull-through operation combined with ostomy closure.After the operation, 1 case had anastomotic leakage and septic shock, 1 case had incision dehiscence, 3 cases had adhesive intestinal obstruction and 1 case occurred left lung consolidation.During a follow-up period of 3-36 months, their recovery condition was acceptable.Conclusions:For those neonates receiving enterostomy, detailed medical history should be enquired before the second stage operation.The combination of barium enema and colonoscopy, 24-hour delayed radiography and multi-point intestinal biopsy with intraoperative exploration conduced to selecting appropriate individualized surgical schemes and reducing surgical complications, thus improving the long-term quality of life of children.

Objective:To explore the application value of 3D printing technology in the precise resection of pediatric retroperitoneal neuroblastoma(NB).Methods:Eleven patients with retroperitoneal NB admitted to the Department of Pediatric Surgery of Henan Provincial People′s Hospital from March 2017 to December 2019 were selected in this study, and all of them received preoperative plain and enhanced computed tomography (CT) scanning of whole abdomen.The original data were imported into the mimics 20 software for processing.The tumor and important peripheral blood vessels were reconstructed with such commands as multi-layer edition, threshold adjustment, regional growth and 3D edition to output the standardized 3D printing stl format file and print out the tumor model by 3D printer.Based on the 3D reconstruction results, the precise surgical planning was performed to determine the optimal operative scheme.The data of all cases were analyzed retrospectively.The parents of these children were investigated by questionnaire and the results were statistically analyzed.Results:All 11 patients successfully underwent the operation under the planning of 3D printing technology.Nine of them had complete tumor resection and 2 cases had palliative resection.The operation time ranged from 140 min to 750 min, with an average of 210 min; the bleeding volume ranged from 100 mL to 1 500 mL, with an average of 240 mL; the postoperative pathological reports revealed that 9 children had NB and 2 cases had ganglioneuroblastoma.There was no case of perioperative death.There were 2 cases of postoperative complications, including 1 case of severe diarrhea with chylous leakage, which was relieved after the treatment of parenteral nutrition and somatostatin, and 1 case of left lower extremity weight-bearing difficulty after left iliac artery anastomosis, which was improved after rehabilitation physical therapy.The follow-up time ranged from 1 month to 30 months.There were 2 cases recurred in the original operation area.One patient received reoperation after chemotherapy; the other continued chemotherapy in progress.As for the 2 cases of palliative resection, they received chemotherapy and survived with tumor.After using 3D model to communicate with children′s parents, their awareness rate of disease, operation mode and postoperative complications were significantly higher than that of CT.Conclusions:3D printing technology can accurately evaluate the structural relationship between retroperitoneal NB and important peripheral blood vessels in children, which contributes to improving the parents′ cognition of the operation mode, and is conducive to selecting the optimal operation scheme and achieving precise tumor resection.

Objective:To explore the value of detecting plasma donor-derived free DNA (dd-cfDNA) fraction in distinguishing antibody mediated-rejection (ABMR) and T cell-mediated rejection (TCMR) of renal allografts.Methods:Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1, 2017 to July 18, 2019 were retrospectively included. Based on pathological classification of Banff renal allograft rejection in 2017, the patients were divided into ABMR group and TCMR group, and the latter was subdivided into TCMR Ⅰ subgroup and TCMR Ⅱ subgroup. The second generation sequencing and target region capture were used to detect candidates' peripheral blood dd-cfDNA. The demographic and clinicopathological data of the two groups were compared. The receiver operating characteristic curve (ROC) was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results:A total of 60 patients with acute rejection of renal transplantation were enrolled in this study, including 42 patients in TCMR group and 18 patients in ABMR group. The plasma dd-cfDNA percentage (%) in the ABMR group was significantly higher than that in the TCMR group [2.33(1.19, 4.30)% vs 0.98(0.50, 1.82)%, P=0.001]. The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group [0.94(0.60, 2.27) ng/ml vs 0.43(0.20, 0.96) ng/ml, P=0.003]. ROC analysis to discriminate TCMR from ABMR showed that, the area under the curve ( AUC) of dd-cfDNA% was 0.76(95% CI 0.64-0.88), when the threshold was 1.11%, the sensitivity and specificity were 88.89% and 59.52%, respectively; the AUC of absolute value of dd-cfDNA was 0.74(95% CI 0.61-0.86), when the threshold was 0.53 ng/ml, the sensitivity was 88.89% and the specificity was 54.76%. TCMR subgroups were further analyzed, there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA (both P>0.05); dd-cfDNA% in ABMR group was apparently higher than that in TCMRⅠ subgroups ( P=0.008) and TCMRⅡsubgroup ( P=0.030). The absolute value of dd-cfDNA in ABMR group was significantly higher than that in TCMRⅠsubgroups ( P=0.003). Conclusion:Plasma dd-cfDNA level may help to distinguish between ABMR and TCMR rejection.

BK polyomavirus-associated nephropathy (BKPyVAN) is a common cause of allograft failure. However, differentiation between BKPyVAN and type I T cell-mediated rejection (TCMR) is challenging when simian virus 40 (SV40) staining is negative, because of the similarities in histopathology. This study investigated whether donor-derived cell-free DNA (ddcfDNA) can be used to differentiate BKPyVAN. Target region capture sequencing was applied to detect the ddcfDNAs of 12 recipients with stable graft function, 22 with type I TCMR, 21 with proven BKPyVAN, and 5 with possible PyVAN. We found that urinary ddcfDNA levels were upregulated in recipients with graft injury, whereas plasma ddcfDNA levels were comparable for all groups. The median urinary concentrations and fractions of ddcfDNA in proven BKPyVAN recipients were significantly higher than those in type I TCMR recipients (10.4 vs. 6.1 ng/mL,